The effect of social deprivation on clinical outcomes and the use of treatments in the UK cystic fibrosis population: a longitudinal study.
Poorer socioeconomic circumstances have been linked with worse outcomes in cystic fibrosis. We assessed whether a relation exists between social deprivation and individual's clinical and health-care outcomes. We did a longitudinal registry study of the UK cystic fibrosis population younger than 40 years (8055 people with 49337 observations for weight, the most commonly collected outcome, between Jan 1, 1996, and Dec 31, 2009). We assessed data for weight, height, body-mass index, percent predicted forced expiratory volume in 1 s (%FEV1), risk of Pseudomonas aeruginosa colonisation, and the use of major cystic fibrosis treatment modalities. We used mixed effects models to assess the association between small-area deprivation and clinical and health-care outcomes, adjusting for clinically important covariates. We give continuous outcomes as mean differences, and binary outcomes as odds ratios, comparing extremes of deprivation quintile. Compared with the least deprived areas, children from the most deprived areas weighed less (standard deviation [SD] score -0·28, 95% CI -0·38 to -0·18), were shorter (-0·31, -0·40 to -0·21, and had a lower body-mass index (-0·13, -0·22 to -0·04), were more likely to have chronic P aeruginosa infection (odds ratio 1·89, 95% CI 1·34 to 2·66), and have a lower %FEV1 (-4·12 percentage points, 95% CI -5·01 to -3·19). These inequalities were apparent very early in life and did not widen thereafter. On a population level, after adjustment for disease severity, children in the most deprived quintile were more likely to receive intravenous antibiotics (odds ratio 2·52, 95% CI 1·92 to 3·17) and nutritional treatments (1·78, 1·44 to 2·20) compared with individuals in the least deprived quintile. Patients from the most disadvantaged areas were less likely to receive DNase or inhaled antibiotic treatment. In the UK, children with cystic fibrosis from more disadvantaged areas have worse growth and lung function compared with children from more affluent areas, but these inequalities do not widen with advancing age. Clinicians consider deprivation status, as well as disease status, when making decisions about treatments, and this might mitigate some effects of social disadvantage. Medical Research Council (UK).


Shifts in Traditional Methods of Coping Among Elderly Bedouin Men.
Elderly Bedouin men in southern Israel are a unique traditional population living in remote unrecognized villages and experiencing rapid social transition, in addition to deep poverty and political tension. In this study, we aimed to explore stressful events, as self-defined by the participants, and the ways in which these men have coped with those stressful events. This study involved 12 men, aged 69⁻74, who participated in in-depth narrative interviews during which they were asked about transformative stressful events in their lives and how they had managed, understood, and utilized human capital, meaning-making, and other methods of coping. Analysis of the interviews revealed several themes: (a) the definition of stressful events within the cultural context, (b) the use of human capital to overcome those events, (c) the transformation of experience from hindsight into a didactic narrative that can be used to assign meaning to past events, which can then be passed on to the next generation, and (d) cultural transition as a catalyst for the creation of new understandings of events. This paper sheds new light on how elderly indigenous Bedouin men self-define stressful situations within a complex and unstable cultural context. This specific context, can help us to gain insight into how indigenous impoverished older men in similar contexts may self-define their stress and coping, based on the types of generalization accepted in qualitative research. The methodological contribution of this work lies in its use of narrative to culturally contextualize phenomenological meaning structures. Its theoretical contribution lies in its examination of the concept of stress within a specific cultural context.


The Effect of Blood on Apical Microleakage of Epiphany and AH26: An In Vitro Study.
Blood contamination of the canal during preparation can be a major problem in endodontics; this may result in apical microleakage. The aim of this in vitro study was to evaluate the effect of blood on apical microleakage of a resin-based root canal sealer (AH26) and a polymer-based root canal sealer (Epiphany). In this experimental study, 50 decoronated central incisors and canine teeth were prepared by RaCe rotary system and randomly divided into 4 groups (n=10). Groups A1 and A2 were obturated by Epiphany/Resilon and AH26/Gutta-percha, respectively. The obturations were performed with a single cone technique after drying root canals. In B1 and B2 groups, the test groups, 0.02cc citrated human blood was injected into dried root canals and they were obturated in the same manner. Ten specimens were served as positive and negative controls (n=5).The apical leakage was measured by means of a computerized fluid filtration method after 1 day and 3 weeks. The data was analyzed by One-Sample Kolmogorov-Smirnov, Independent Sample t-test and univariate analysis. Statistical significances were preset at α=0.05. There was no significant difference in apical microleakage of the two sealers after 1 day and 3 weeks in dry and blood environment (P>0.05). Sealer and environment had no interaction (P>0.05). Blood contamination has no significant effect on the apical microleakage of Epiphny and AH26.


Mammalian gastrointestinal tract parameters modulating the integrity, surface properties, and absorption of food-relevant nanomaterials.
Many natural chemicals in food are in the nanometer size range, and the selective uptake of nutrients with nanoscale dimensions by the gastrointestinal (GI) tract is a normal physiological process. Novel engineered nanomaterials (NMs) can bring various benefits to food, e.g., enhancing nutrition. Assessing potential risks requires an understanding of the stability of these entities in the GI lumen, and an understanding of whether or not they can be absorbed and thus become systemically available. Data are emerging on the mammalian in vivo absorption of engineered NMs composed of chemicals with a range of properties, including metal, mineral, biochemical macromolecules, and lipid-based entities. In vitro and in silico fluid incubation data has also provided some evidence of changes in particle stability, aggregation, and surface properties following interaction with luminal factors present in the GI tract. The variables include physical forces, osmotic concentration, pH, digestive enzymes, other food, and endogenous biochemicals, and commensal microbes. Further research is required to fill remaining data gaps on the effects of these parameters on NM integrity, physicochemical properties, and GI absorption. Knowledge of the most influential luminal parameters will be essential when developing models of the GI tract to quantify the percent absorption of food-relevant engineered NMs for risk assessment.


Brain amino acid abnormalities in liver disease--a postmortem study.
In a postmortem exploratory study, we examined whether specific amino acid abnormalities associated with liver diseases in vivo may also be detected in human brain samples obtained at clinical autopsies. The branched-chain amino acids (BCAA: valine, leucine, isoleucine) were decreased in the group of patients with liver diseases compared with the control group, whereas the aromatic amino acids (AAA: phenylalanine, tyrosine) were increased. However, the ranges overlapped significantly and were not statistically different. The molar ratio BCAA/AAA was determined to be 1.92 in the collection of patients with liver diseases compared with 2.27 in the control group. In patients with liver disease, ornithine concentrations in the brain appeared significantly decreased whereas glutamine was significantly increased. No significant difference was found in the brain concentrations of proline. Amino acid analysis may contribute to the understanding of pathophysiological mechanisms of liver disease, which are discussed, and may supplement the postmortem diagnosis.


Guanine nucleotide and cation regulation of the binding of [3H]cyclohexyladenosine and [3H]diethylphenylxanthine to adenosine A1 receptors in brain membranes.
Guanine nucleotides, divalent cations, and sodium differentially regulate agonist and antagonist binding to adenosine A1 receptors in brain membranes. Guanine nucleotides decrease the binding of the adenosine A1 receptor agonist [3H]N6-cyclohexyladenosine ([3H]CHA) to guinea pig and bovine brain membranes by about 50% at 1--3 microM, while not affecting binding of the antagonist [3H]1,3-diethyl-8-phenylxanthine ([3H]DPX) to A1 receptors in bovine brain. GTP decreases the potency of agonists competing for [3H]DPX binding by 3--6 times, without altering the potency of antagonists. This effect can be used to grade experimental substances along an adenosine agonist-antagonist continuum. The 66% inhibition of [3H]CHA binding by 1 mM EDTA, with no change in [3H]DPX binding, suggests that endogenous divalent cations may regulate adenosine receptor interactions. Removal of endogenous divalent cations by EDTA treatment greatly increases the enhancement of [3H]CHA binding by divalent cations. Specific binding of [3H]CHA to guinea pig brain is increased 150--170% by 0.3--1.0 mM Mn2+, Mg2+, and Ca2+ following EDTA preincubation, secondary to an increase in apparent affinity and receptor number. Sodium ions also selectively regulate the binding of [3H]CHA. Sodium decreases [3H]CHA binding 40%, whereas lithium and potassium are ineffective. Sodium does not affect [3H]DPX binding.


Tako-Tsubo-like cardiomyopathy: specific ECG findings, characterization and clinical findings in a European single center.
A newly discovered heart syndrome mimicking acute coronary syndrome has been termed 'Tako-Tsubo cardiomyopathy' (TTC). Differentiation from acute myocardial infarction using the ECG is an important issue in clinical practice. We retrospectively analyzed patients admitted for cardiac catheterization between September 2003 and September 2006. From 26,593 cardiac catheterization procedures, we identified 21 patients with suggested TTC (0.08%). Trigger mechanisms were present in 38.1%; all patients had elevated troponin levels (mean 3.9 ng/ml). Median age was 68.4 years; 90.5% were female. Hypertension was seen in 85.7% and atrial fibrillation in 19.1%. Specific ECG findings related to a TTC are: a mild elevation of the ST segment arising from the S curve of the QRS complex, where the maximum ST segment elevation at the basis of the T wave is <1.5 mm, T-wave inversion, absence of ST segment depression and a summated amplitude of the S curve in V1 plus R in V6 <1.5 mV. An intraventricular gradient was seen in 9.5%; coronary atherosclerosis was detected in 57.1%. Follow-Up Data: Mean follow-up was 13.2 months. 47.6% were free from angina or dyspnea, most of the patients received beta-blockers/ACE inhibitors (76.2%). One patient had a sudden cardiac death (4.8%), 1 patient became an implantable cardioverter-defibrillator primarily due to resuscitation. TTC is a rare syndrome mimicking acute coronary syndrome that shows a specific ECG pattern and does not appear to be an unambiguously benign disease.


Net charge per residue modulates conformational ensembles of intrinsically disordered proteins.
Intrinsically disordered proteins (IDPs) adopt heterogeneous ensembles of conformations under physiological conditions. Understanding the relationship between amino acid sequence and conformational ensembles of IDPs can help clarify the role of disorder in physiological function. Recent studies revealed that polar IDPs favor collapsed ensembles in water despite the absence of hydrophobic groups--a result that holds for polypeptide backbones as well. By studying highly charged polypeptides, a different archetype of IDPs, we assess how charge content modulates the intrinsic preference of polypeptide backbones for collapsed structures. We characterized conformational ensembles for a set of protamines in aqueous milieus using molecular simulations and fluorescence measurements. Protamines are arginine-rich IDPs involved in the condensation of chromatin during spermatogenesis. Simulations based on the ABSINTH implicit solvation model predict the existence of a globule-to-coil transition, with net charge per residue serving as the discriminating order parameter. The transition is supported by quantitative agreement between simulation and experiment. Local conformational preferences partially explain the observed trends of polymeric properties. Our results lead to the proposal of a schematic protein phase diagram that should enable prediction of polymeric attributes for IDP conformational ensembles using easily calculated physicochemical properties of amino acid sequences. Although sequence composition allows the prediction of polymeric properties, interresidue contact preferences of protamines with similar polymeric attributes suggest that certain details of conformational ensembles depend on the sequence. This provides a plausible mechanism for specificity in the functions of IDPs.


Proteome-wide mapping of immune features onto Plasmodium protein three-dimensional structures.
Humoral immune responses against the malaria parasite are an important component of a protective immune response. Antibodies are often directed towards conformational epitopes, and the native structure of the antigenic region is usually critical for antibody recognition. We examined the structural features of various Plasmodium antigens that may impact on epitope location, by performing a comprehensive analysis of known and modelled structures from P. falciparum. Examining the location of known polymorphisms over all available structures, we observed a strong propensity for polymorphic residues to be exposed on the surface and to occur in particular secondary structure segments such as hydrogen-bonded turns. We also utilised established prediction algorithms for B-cell epitopes and MHC class II binding peptides, examining predicted epitopes in relation to known polymorphic sites within structured regions. Finally, we used the available structures to examine polymorphic hotspots and Tajima's D values using a spatial averaging approach. We identified a region of PfAMA1 involving both domains II and III under a high degree of balancing selection relative to the rest of the protein. In summary, we developed general methods for examining how sequence-based features relate to one another in three-dimensional space and applied these methods to key P. falciparum antigens.


Contextual influences on the perception of pregnant women who use drugs: Information about women's childhood trauma history reduces punitive attitudes.
Punitive attitudes and consequences (e.g., incarceration) for prenatal illicit drug use persist in the United States despite evidence that these policies are ineffective and even harmful to women and children. For instance, the threat of these consequences can deter women from seeking healthcare, prenatal care, and drug treatment. Punitive responses may persist due to pejorative public perceptions of pregnant women who use illicit drugs. Although there is evidence that contextual information about prenatal drug use (e.g., drug type) can change such perceptions, other contextual influences are unknown. This experimental study tested whether receiving contextual information about a pregnant woman who uses drugs (specifically, her childhood trauma history) reduces punitive and increases supportive attitudes toward the woman. In a vignette-based 2(pregnancy status: pregnant/not pregnant) x 2(history of childhood trauma: interpersonal/non-interpersonal) between-subjects design, young adult university participants (N = 461) were randomly assigned to read a vignette about a woman who uses methamphetamine. Punitive attitudes were significantly reduced by information that the pregnant woman had a history of childhood trauma, especially interpersonal (versus non-interpersonal) trauma (ηp2 = .115). Supportive attitudes were not impacted (ηp2 = .005). Information about the pregnant woman's trauma history predicted less agreement with incarcerating her, only indirectly, through less punitive attitudes (R2 = .21). Reductions in punitive attitudes were on the order of 1.5-2 points on 5-point self-report scales and controlled for participant gender and political conservatism. Results have practical implications for interdisciplinary work aimed at unlocking greater support for policies that help pregnant women make safe, informed decisions with dignity and access to healthcare.


Frailty as a predictor of the incidence and course of depressed mood.
Late-life depression and physical frailty are supposed to be reciprocally associated, however, longitudinal studies are lacking. This study examines whether physical frailty predicts a higher incidence of depression, as well as a less favorable course of depression. A population-based cohort study of 888 people aged 65 years and over with follow-up measures at 3, 6, and 9 years. Cox proportional hazards models adjusted for age, sex, education, smoking, alcohol usage, and global cognitive functioning were applied to calculate the incidence of depressed mood in those nondepressed at baseline (n = 699) and remission in those with depressed mood at baseline (n = 189). Depressed mood onset or remission was defined as crossing the cut-off score of 20 points on the Center for Epidemiological Studies-Depression Scale combined with a relevant change in this score. Physical frailty was based on the presence of ≥ 3 out of 5 components (ie, weight loss, weakness, slowness, exhaustion, and low physical activity level). A total of 214 out of 699 (30.6%) nondepressed persons developed depressed mood during follow-up. Physical frailty predicted the onset of depressed mood with a hazard rate of 1.26 (95% confidence interval 1.09-1.45, P = .002). Of the 189 persons with depressed mood at baseline, 96 (50.8%) experienced remission during follow-up. Remission was less likely in the presence of a higher level of physical frailty (hazard rate = 0.72, 95% confidence interval 0.58-0.91, P = .005). Because physical frailty predicts both the onset and course of late-life depressed mood, physical frailty should receive more attention in mental health care planning for older persons as well as its interference with treatment. Future studies into the pathophysiological mechanisms may guide the development of new treatment opportunities for these vulnerable patients.


Conservation and early expression of zebrafish tyrosine kinases support the utility of zebrafish as a model for tyrosine kinase biology.
Tyrosine kinases have significant roles in cell growth, apoptosis, development, and disease. To explore the use of zebrafish as a vertebrate model for tyrosine kinase signaling and to better understand their roles, we have identified all of the tyrosine kinases encoded in the zebrafish genome and quantified RNA expression of selected tyrosine kinases during early development. Using profile hidden Markov model analysis, we identified 122 zebrafish tyrosine kinase genes and proposed unambiguous gene names where needed. We found them to be organized into 39 nonreceptor and 83 receptor type, and 30 families consistent with human tyrosine kinase family assignments. We found five human tyrosine kinase genes (epha1, bmx, fgr, srm, and insrr) with no identifiable zebrafish ortholog, and one zebrafish gene (yrk) with no identifiable human ortholog. We also found that receptor tyrosine kinase genes were duplicated more often than nonreceptor tyrosine kinase genes in zebrafish. We profiled expression levels of 30 tyrosine kinases representing all families using direct digital detection at different stages during the first 24 hours of development. The profiling experiments clearly indicate regulated expression of tyrosine kinases in the zebrafish, suggesting their role during early embryonic development. In summary, our study has resulted in the first comprehensive description of the zebrafish tyrosine kinome.


Psychological management of post-stroke depression.
To evaluate existing research literature on psychological interventions to manage post-stroke depression (PSD). First, the particular nature of post-stroke depression (PSD) was established. Then, relevant published material was identified through computerized literature searches (to May 1999) and via the first author's clinical experience in the area. This material was critically reviewed against recommended standards for the empirical validation of treatment effectiveness. Methodological limitations in existing research prevent a conclusion as any one psychological intervention has empirical support for its effectiveness. However, cognitive behaviour therapy in particular was identified as worthy of further investigation. It is recommended that future research investigates treatments for PSD appropriate for those with cognitive impairment and communication difficulties, younger versus older stroke victims and those in institutional settings. Studies should also consider the necessity of specialist assessment strategies and allow for possible subtypes of PSD for which psychological treatments might be differentially effective.


The alkaline cleavage and borohydride reduction of cartilage proteoglycan.
A method for the rapid isolation and purification of proteoglycan by using neutral solutions of LiBr for extraction and density-gradient centrifugation is described. The effect of 0.5m-KOH on isolated proteoglycan has been studied by using NaB(3)H(4) to reduce and label the chondroitin sulphate chains released. This study has established: (a) that at least 95% of the chondroitin sulphate chains are attached to the proteoglycan by alkali-labile bonds between xylose and serine; (b) that random degradation of the chondroitin sulphate chains does not occur to any significant extent; (c) that the method is convenient for the determination of polysaccharide number-average molecular weights.


Evasion of IFN-γ signaling by Francisella novicida is dependent upon Francisella outer membrane protein C.
Francisella tularensis is a Gram-negative facultative intracellular bacterium and the causative agent of the lethal disease tularemia. An outer membrane protein (FTT0918) of F. tularensis subsp. tularensis has been identified as a virulence factor. We generated a F. novicida (F. tularensis subsp. novicida) FTN_0444 (homolog of FTT0918) fopC mutant to study the virulence-associated mechanism(s) of FTT0918. The ΔfopC strain phenotype was characterized using immunological and biochemical assays. Attenuated virulence via the pulmonary route in wildtype C57BL/6 and BALB/c mice, as well as in knockout (KO) mice, including MHC I, MHC II, and µmT (B cell deficient), but not in IFN-γ or IFN-γR KO mice was observed. Primary bone marrow derived macrophages (BMDM) prepared from C57BL/6 mice treated with rIFN-γ exhibited greater inhibition of intracellular ΔfopC than wildtype U112 strain replication; whereas, IFN-γR KO macrophages showed no IFN-γ-dependent inhibition of ΔfopC replication. Moreover, phosphorylation of STAT1 was downregulated by the wildtype strain, but not the fopC mutant, in rIFN-γ treated macrophages. Addition of NG-monomethyl-L-arginine, an NOS inhibitor, led to an increase of ΔfopC replication to that seen in the BMDM unstimulated with rIFN-γ. Enzymatic screening of ΔfopC revealed aberrant acid phosphatase activity and localization. Furthermore, a greater abundance of different proteins in the culture supernatants of ΔfopC than that in the wildtype U112 strain was observed. F. novicida FopC protein facilitates evasion of IFN-γ-mediated immune defense(s) by down-regulation of STAT1 phosphorylation and nitric oxide production, thereby promoting virulence. Additionally, the FopC protein also may play a role in maintaining outer membrane stability (integrity) facilitating the activity and localization of acid phosphatases and other F. novicida cell components.


Muscular outputs during dynamic bench press under stable versus unstable conditions.
Previous studies have suggested that resistance training exercise under unstable conditions decreases the isometric force output, yet little is known about its influence on muscular outputs during dynamic movement. The objective of this study was to investigate the effect of an unstable condition on power, force, and velocity outputs during the bench press. Twenty male collegiate athletes (mean age, 21.3 +/- 1.5 years; mean height, 167.7 +/- 7.7 cm; mean weight, 75.9 +/- 17.5 kg) participated in this study. Each subject attempted 3 sets of single bench presses with 50% of 1 repetition maximum (1RM) under a stable condition with a flat bench and an unstable condition with a Swiss ball. Acceleration data were obtained with an accelerometer attached to the center of a barbell shaft, and peak outputs of power, force, and velocity were computed. Although significant loss of the peak outputs was found under the unstable condition (p < 0.017), their reduction rates remained relatively low, approximately 6% for force and 10% for power and velocity outputs, compared with previous findings. Such small reduction rates of muscular outputs may not compromise the training effect. Prospective studies are necessary to confirm whether the resistance training under an unstable condition permits the improvement of dynamic performance and trunk stability.


Glucagon and Amino Acids Are Linked in a Mutual Feedback Cycle: The Liver-α-Cell Axis.
Glucagon is usually viewed as an important counterregulatory hormone in glucose metabolism, with actions opposing those of insulin. Evidence exists that shows glucagon is important for minute-to-minute regulation of postprandial hepatic glucose production, although conditions of glucagon excess or deficiency do not cause changes compatible with this view. In patients with glucagon-producing tumors (glucagonomas), the most conspicuous signs are skin lesions (necrolytic migratory erythema), while in subjects with inactivating mutations of the glucagon receptor, pancreatic swelling may be the first sign; neither condition is necessarily associated with disturbed glucose metabolism. In glucagonoma patients, amino acid turnover and ureagenesis are greatly accelerated, and low plasma amino acid levels are probably at least partly responsible for the necrolytic migratory erythema, which resolves after amino acid administration. In patients with receptor mutations (and in knockout mice), pancreatic swelling is due to α-cell hyperplasia with gross hypersecretion of glucagon, which according to recent groundbreaking research may result from elevated amino acid levels. Additionally, solid evidence indicates that ureagenesis, and thereby amino acid levels, is critically controlled by glucagon. Together, this constitutes a complete endocrine system; feedback regulation involving amino acids regulates α-cell function and secretion, while glucagon, in turn, regulates amino acid turnover.


Multiple versus single injections of fluorescent microspheres for the determination of regional organ blood flow in septic sheep.
Determination of regional blood flow by the injection of microspheres in sepsis models is crucial for the experimental evaluation of the influence of experimental treatment strategies on organ perfusion. However, multiple injections may critically increase the total quantity of microspheres, thereby restricting regional microcirculation and altering the results of blood flow measurements. This study was designed to compare the results of multiple versus single injections of microspheres in an established ovine sepsis model. Injury was induced by smoke inhalation and instillation of Pseudomonas aeruginosa into the lungs. Twenty sheep were studied for 4, 8, 12, 18, or 24 h, respectively. Microspheres were injected at the end of the study period and the animals were euthanized and organ tissues were harvested. Another four sheep were studied for 24 h and multiple microsphere injections were performed at the above indicated time points in the same animals. Tracheal blood flow significantly increased and blood flow to the pancreas and ileum significantly decreased versus baseline in both groups (P < 0.05 each). Blood flow to the ileum, renal cortex and skin did not significantly change versus baseline in both groups (P > 0.05). Blood flow was higher to the trachea in the multiple injection group at 18 h (P = 0.048) and to the ileum at 12 h (P = 0.049), and lower to the skin at 18 h (P = 0.015). In conclusion, the results indicate that multiple versus single microsphere injections induced no or negligible alterations during ovine sepsis. This finding may help reduce the quantity of animals needed in future experiments.


[Study of intralysosomal protein catabolism using lysosomotropic agents--proteolysis and protease inhibitors].
Inhibition of intralysosomal catabolism of proteins was studied in rat liver cells using lysosomotropic drugs suramin (single administration at a dose of 250 mg/kg within 24 and 48 hrs) and chloroquine (at a dose of 30 mg/100 g within 0.5, 1, 3, 6 and 12 hrs after administration). Suramine inhibited pure preparations of cathepsin B and L, while chloroquine inhibited cathepsins B, L and H. The inhibitors were effective in vitro at lower concentrations as compared with these administered in vivo. Less distinct inhibitory effect was observed in the incubation mixture containing an extract of rat liver lysosomes. After administration of the inhibitors in vivo the activity of lysosomal proteinases was not inhibited in rat liver homogenates, which appears to occur due to complex formation between these lysosomotropic drugs and proteins.


Cold storage of rat hepatocyte suspensions for one week in a customized cold storage solution--preservation of cell attachment and metabolism.
Primary hepatocytes are of great importance for basic research as well as cell transplantation. However, their stability, especially in suspension, is very low. This feature severely compromises storage and shipment. Based on previous studies with adherent cells, we here assessed cold storage injury in rat hepatocyte suspensions and aimed to find a cold storage solution that preserves viability, attachment ability and functionality of these cells. Rat hepatocyte suspensions were stored in cell culture medium, organ preservation solutions and modified TiProtec solutions at 4°C for one week. Viability and cell volume were determined by flow cytometry. Thereafter, cells were seeded and density and metabolic capacity (reductive metabolism, forskolin-induced glucose release, urea production) of adherent cells were assessed. Cold storage injury in hepatocyte suspensions became evident as cell death occurring during cold storage or rewarming or as loss of attachment ability. Cell death during cold storage was not dependent on cell swelling and was almost completely inhibited in the presence of glycine and L-alanine. Cell attachment could be greatly improved by use of chloride-poor solutions and addition of iron chelators. Using a chloride-poor, potassium-rich storage solution containing glycine, alanine and iron chelators, cultures with 75% of the density of control cultures and with practically normal cell metabolism could be obtained after one week of cold storage. In the solution presented here, cold storage injury of hepatocyte suspensions, differing from that of adherent hepatocytes, was effectively inhibited. The components which acted on the different injurious processes were identified.


A web-based Delphi study on the indications of chest radiographs for patients in ICUs.
Strategies for ordering bedside chest radiographs (CXRs) have substantial logistic and financial consequences in the ICU. Many of the indications for CXRs in the ICU are controversial, such as the ordering of daily routine CXRs for intubated patients. The opinions of intensivists about ordering CXRs have not been reported. Comparing these opinions to established guidelines and identifying situations where opinions diverge in the absence of guidelines are of considerable interest. We asked 190 intensivists from 34 ICUs in the area of Paris, France, to anonymously complete a 29-item questionnaire about their opinions regarding the ordering of CXRs; each item described a clinical scenario. Of the 29 scenarios, 10 dealt with the placement of medical devices, 8 with the presence of medical devices, and 11 with other clinical situations. The study was based on a Delphi process deployed over the Internet through an original software application. Three Delphi rounds were run between January and March 2006, using the same questionnaire. Detailed feedback for the answers given during the previous round was supplied to each intensivist solicited for updating his answers. Eighty-two intensivists from 32 ICUs completed the study. A consensus emerged that routine CXRs were necessary for eight scenarios and unnecessary for two scenarios. The study also shed light on items without a consensus. In particular, 75% of intensivists (58% on the first round) did not support obtaining daily routine CXRs in intubated patients. The study underlines situations in which intensivists do not support the guidelines and outlines recommendations likely to be followed in clinical practice.


Blood-wall fluttering instability as a physiomarker of the progression of thoracic aortic aneurysms.
The diagnosis of aneurysms is informed by empirically tracking their size and growth rate. Here, by analysing the growth of aortic aneurysms from first principles via linear stability analysis of flow through an elastic blood vessel, we show that abnormal aortic dilatation is associated with a transition from stable flow to unstable aortic fluttering. This transition to instability can be described by the critical threshold for a dimensionless number that depends on blood pressure, the size of the aorta, and the shear stress and stiffness of the aortic wall. By analysing data from four-dimensional flow magnetic resonance imaging for 117 patients who had undergone cardiothoracic imaging and for 100 healthy volunteers, we show that the dimensionless number is a physiomarker for the growth of thoracic ascending aortic aneurysms and that it can be used to accurately discriminate abnormal versus natural growth. Further characterization of the transition to blood-wall fluttering instability may aid the understanding of the mechanisms underlying aneurysm progression in patients.


Protocol for the process evaluation of a cluster randomised controlled trial to determine the effectiveness and cost-effectiveness of independent pharmacist prescribing in care home: the CHIPPS study.
Prescribing, monitoring and administration of medicines in care homes could be improved. A cluster randomised controlled trial (RCT) is ongoing to evaluate the effectiveness of an independent prescribing pharmacist assuming responsibility for medicines management in care homes compared to usual care. To conduct a mixed-methods process evaluation of the RCT, in line with Medical Research Council (MRC) process evaluation guidance, to inform interpretation of main trial findings and if the service is found to be effective and efficient, to inform subsequent implementation. 1. To describe the intervention as delivered in terms of quality, quantity, adaptations and variations across triads and time. 2. To explore the effects of individual intervention components on the primary outcomes. 3. To investigate the mechanisms of impact. 4. To describe the perceived effectiveness of relevant intervention components [including pharmacist independent prescriber (PIP) training and care home staff training] from participant [general practitioner (GP), care home, PIP and resident/relative] perspectives. 5. To describe the characteristics of GP, care home, PIP and resident participants to assess reach. 6. To estimate the extent to which intervention delivery is normalised among the intervention healthcare professionals and related practice staff. A mix of quantitative (surveys, record reviews) and qualitative (interviews) approaches will be used to collect data on the extent of the delivery of detailed tasks required to implement the new service, to collect data to confirm the mechanism of impact as hypothesised in the logic model, to collect explanatory process and final outcome data, and data on contextual factors which could have facilitated or hindered effective and efficient delivery of the service. Recruitment is ongoing and the trial should complete in early 2020. The systematic and comprehensive approach that is being adopted will ensure data is captured on all aspects of the study, and allow a full understanding of the implementation of the service and the RCT findings. With so many interrelated factors involved it is important that a process evaluation is undertaken to enable us to identify which elements of the service were deemed to be effective, explain any differences seen, and identify enablers, barriers and future adaptions. ISRCTN17847169. Date registered: 15 December 2017.


Cost-Effectiveness of Tafamidis Therapy for Transthyretin Amyloid Cardiomyopathy.
In patients with transthyretin amyloid cardiomyopathy, tafamidis reduces all-cause mortality and cardiovascular hospitalizations and slows decline in quality of life compared with placebo. In May 2019, tafamidis received expedited approval from the US Food and Drug Administration as a breakthrough drug for a rare disease. However, at $225 000 per year, it is the most expensive cardiovascular drug ever launched in the United States, and its long-term cost-effectiveness and budget impact are uncertain. We therefore aimed to estimate the cost-effectiveness of tafamidis and its potential effect on US health care spending. We developed a Markov model of patients with wild-type or variant transthyretin amyloid cardiomyopathy and heart failure (mean age, 74.5 years) using inputs from the ATTR-ACT trial (Transthyretin Amyloidosis Cardiomyopathy Clinical Trial), published literature, US Food and Drug Administration review documents, healthcare claims, and national survey data. We compared no disease-specific treatment ("usual care") with tafamidis therapy. The model reproduced 30-month survival, quality of life, and cardiovascular hospitalization rates observed in ATTR-ACT; future projections used a parametric survival model in the control arm, with constant hazards reduction in the tafamidis arm. We discounted future costs and quality-adjusted life-years by 3% annually and examined key parameter uncertainty using deterministic and probabilistic sensitivity analyses. The main outcomes were lifetime incremental cost-effectiveness ratio and annual budget impact, assessed from the US healthcare sector perspective. This study was independent of the ATTR-ACT trial sponsor. Compared with usual care, tafamidis was projected to add 1.29 (95% uncertainty interval, 0.47-1.75) quality-adjusted life-years at an incremental cost of $1 135 000 (872 000-1 377 000), resulting in an incremental cost-effectiveness ratio of $880 000 (697 000-1 564 000) per quality-adjusted life-year gained. Assuming a threshold of $100 000 per quality-adjusted life-year gained and current drug price, tafamidis was cost-effective in 0% of 10 000 probabilistic simulations. A 92.6% price reduction from $225 000 to $16 563 would be necessary to make tafamidis cost-effective at $100 000/quality-adjusted life-year. Results were sensitive to assumptions related to long-term effectiveness of tafamidis. Treating all eligible patients with transthyretin amyloid cardiomyopathy in the United States with tafamidis (n=120 000) was estimated to increase annual healthcare spending by $32.3 billion. Treatment with tafamidis is projected to produce substantial clinical benefit but would greatly exceed conventional cost-effectiveness thresholds at the current US list price. On the basis of recent US experience with high-cost cardiovascular medications, access to and uptake of this effective therapy may be limited unless there is a large reduction in drug costs.


A common toxin fold mediates microbial attack and plant defense.
Many plant pathogens secrete toxins that enhance microbial virulence by killing host cells. Usually, these toxins are produced by particular microbial taxa, such as bacteria or fungi. In contrast, many bacterial, fungal and oomycete species produce necrosis and ethylene-inducing peptide 1 (Nep1)-like proteins (NLPs) that trigger leaf necrosis and immunity-associated responses in various plants. We have determined the crystal structure of an NLP from the phytopathogenic oomycete Pythium aphanidermatum to 1.35A resolution. The protein fold exhibits structural similarities to cytolytic toxins produced by marine organisms (actinoporins). Computational modeling of the 3-dimensional structure of NLPs from another oomycete, Phytophthora parasitica, and from the phytopathogenic bacterium, Pectobacterium carotovorum, revealed a high extent of fold conservation. Expression of the 2 oomycete NLPs in an nlp-deficient P. carotovorum strain restored bacterial virulence, suggesting that NLPs of prokaryotic and eukaryotic origins are orthologous proteins. NLP mutant protein analyses revealed that identical structural properties were required to cause plasma membrane permeabilization and cytolysis in plant cells, as well as to restore bacterial virulence. In sum, NLPs are conserved virulence factors whose taxonomic distribution is exceptional for microbial phytotoxins, and that contribute to host infection by plasma membrane destruction and cytolysis. We further show that NLP-mediated phytotoxicity and plant defense gene expression share identical fold requirements, suggesting that toxin-mediated interference with host integrity triggers plant immunity-associated responses. Phytotoxin-induced cellular damage-associated activation of plant defenses is reminiscent of microbial toxin-induced inflammasome activation in vertebrates and may thus constitute another conserved element in animal and plant innate immunity.


Long-Term Nontoxic Support of Animal Life with Algae.
One 40-g male albino mouse was kept in good health for 66 days in a chamber containing an algal photosynthetic gas exchanger. Carbon dioxide was well controlled, oxygen was slowly increased, and nitrogen was decreased in the chamber. Photosynthesis can support life in a hostile environment for extended periods of time.


Functioning and disability in persons with epilepsy.
This study aimed to use the International Classification of Functioning, Disability, and Health (ICF) to describe the functioning of people with epilepsy. The main questions we asked were as follows: what are the difficulties that people with epilepsy face on the body level and in daily activities? What are the most relevant environmental factors needed to describe disability in patients with epilepsy? Data collected using the ICF checklist are presented. Forty-nine patients were interviewed. In total, 63 categories, of which 30 were from the component of activities and participation, 18 were from body functions and body structures, and 14 were from environmental factors, were identified. Among body functions, sensation of pain, seeing, and emotional functions were reported most frequently. Among activities and participation, several difficulties were reported, particularly when capacity qualifiers and broad differences between capacity and performance were taken into account. Finally, environmental factor categories were mostly reported as facilitators, particularly those connected with health care. The description of a profile of functioning and disability can be more comprehensive with the use of the framework of the ICF, which provides a broad picture of functioning. A deeper understanding of functioning can be gained using the ICF conceptualization of disability because it incorporates all relevant aspects of functioning at the levels of the body and the person, as well as the impact of the environment.


The effect of vaginal bleeding during pregnancy on preterm and small-for-gestational-age births: US National Maternal and Infant Health Survey, 1988.
We examined the effects of vaginal bleeding during pregnancy on preterm and small-for-gestational-age (SGA) births using data from the 1988 US National Maternal and Infant Health Survey. We examined the severity of vaginal bleeding and separated the preterm births into subsets by the degree of prematurity (< 32, 32-33, 34-36 weeks' gestation). We also evaluated associations stratified by race. Multiple logistic regression analysis showed that vaginal bleeding was associated with an increased risk of preterm birth, with a more pronounced elevated risk for preterm birth before 34 weeks' gestation and a notably stronger association for more severe bleeding that occurred in both the first and the second half of pregnancy. Odds ratios, but not risk differences, for birth before 34 weeks' gestation were greater for white women with vaginal bleeding than for black women. There was no association between vaginal bleeding during pregnancy and SGA births.


Ultrastructural observations on sperm storage in the ovary of the platyfish, Xiphophorus maculatus (Teleostei: poeciliidae): the role of the duct epithelium.
Sperm storage is an important phenomenon occurring in viviparous and oviparous teleosts and contributes to the reproductive life history of these forms. There is a paucity of morphological studies on sperm storage in fishes. The majority of these have been confined to the light level of investigation. In this report, we describe, at the ultrastructural level, sperm storage in the viviparous platyfish, Xiphophorus maculatus. Female platyfish, as is typical of the poeciliids, are capable of storing viable sperm for up to several months within the ovary and gonoduct. We observed that sperm stored within inseminated platyfish became associated with specific epithelial cells (SACs) lining the oviduct. Two forms of association were seen: 1) sperm were found within deep surface pits and pockets, and 2) the spermatozoa were taken up and incorporated within the cytoplasm of the SACs. Junctional complexes in the form of tight junctions (zona occludens) and desmosomes at the apico-lateral surfaces of the SACs were prevalent. The junctions could have contributed to allograft survival of the haploid heterogenetic sperm cells within the female's reproductive tract. Our results shed light on the mechanism of sperm storage in the platyfish and could serve as a model for other poeciliid species and teleosts that are capable of storing sperm.


Registration of renal SPECT and 2.5D US images.
Image registration is the process of transforming different image data sets of an object into the same coordinate system. This is a relevant task in the field of medical imaging; one of its objectives is to combine information from different imaging modalities. The main goal of this study is the registration of renal SPECT (Single Photon Emission Computerized Tomography) images and a sparse set of ultrasound slices (2.5D US), combining functional and anatomical information. Registration is performed after kidney segmentation in both image types. The SPECT segmentation is achieved using a deformable model based on a simplex mesh. The 2.5D US image segmentation is carried out in each of the 2D slices employing a deformable contour and Gabor filters to capture multi-scale image features. Moreover, a renal medulla detection method was developed to improve the US segmentation. A nonlinear optimization algorithm is used for the registration. In this process, movements caused by patient breathing during US image acquisition are also corrected. Only a few reports describe registration between SPECT images and a sparse set of US slices of the kidney, and they usually employ an optical localizer, unlike our method, that performs movement correction using information only from the SPECT and US images. Moreover, it does not require simultaneous acquisition of both image types. The registration method and both segmentations were evaluated separately. The SPECT segmentation was evaluated qualitatively by medical experts, obtaining a score of 5 over a scale from 1 to 5, where 5 represents a perfect segmentation. The 2.5D US segmentation was evaluated quantitatively, by comparing our method with an expert manual segmentation, and obtaining an average error of 3.3mm. The registration was evaluated quantitatively and qualitatively. Quantitatively the distance between the manual segmentation of the US images and the model extracted from the SPECT image was measured, obtaining an average distance of 1.07 pixels on 7 exams. The qualitative evaluation was carried out by a group of physicians who assessed the perceived clinical usefulness of the image registration, rating each registration on a scale from 1 to 5. The average score obtained was 4.1, i.e. relevantly useful for medical purposes.


Epigenetic regulator Stuxnet modulates octopamine effect on sleep through a Stuxnet-Polycomb-Octβ2R cascade.
Sleep homeostasis is crucial for sleep regulation. The role of epigenetic regulation in sleep homeostasis is unestablished. Previous studies showed that octopamine is important for sleep homeostasis. However, the regulatory mechanism of octopamine reception in sleep is unknown. In this study, we identify an epigenetic regulatory cascade (Stuxnet-Polycomb-Octβ2R) that modulates the octopamine receptor in Drosophila. We demonstrate that stuxnet positively regulates Octβ2R through repression of Polycomb in the ellipsoid body of the adult fly brain and that Octβ2R is one of the major receptors mediating octopamine function in sleep homeostasis. In response to octopamine, Octβ2R transcription is inhibited as a result of stuxnet downregulation. This feedback through the Stuxnet-Polycomb-Octβ2R cascade is crucial for sleep homeostasis regulation. This study demonstrates a Stuxnet-Polycomb-Octβ2R-mediated epigenetic regulatory mechanism for octopamine reception, thus providing an example of epigenetic regulation of sleep homeostasis.


Population-Level Risk Factors Related to Measles Case Fatality: A Conceptual Framework Based on Expert Consultation and Literature Review.
A better understanding of population-level factors related to measles case fatality is needed to estimate measles mortality burden and impact of interventions such as vaccination. This study aimed to develop a conceptual framework of mechanisms associated with measles case fatality ratios (CFRs) and assess the scope of evidence available for related indicators. Using expert consultation, we developed a conceptual framework of mechanisms associated with measles CFR and identified population-level indicators potentially associated with each mechanism. We conducted a literature review by searching PubMed on 31 October 2021 to determine the scope of evidence for the expert-identified indicators. Studies were included if they contained evidence of an association between an indicator and CFR and were excluded if they were from non-human studies or reported non-original data. Included studies were assessed for study quality. Expert consultation identified five mechanisms in a conceptual framework of factors related to measles CFR. We identified 3772 studies for review and found 49 studies showing at least one significant association with CFR for 15 indicators (average household size, educational attainment, first- and second-dose coverage of measles-containing vaccine, human immunodeficiency virus prevalence, level of health care available, stunting prevalence, surrounding conflict, travel time to major city or settlement, travel time to nearest health care facility, under-five mortality rate, underweight prevalence, vitamin A deficiency prevalence, vitamin A treatment, and general malnutrition) and only non-significant associations for five indicators (antibiotic use for measles-related pneumonia, malaria prevalence, percent living in urban settings, pneumococcal conjugate vaccination coverage, vitamin A supplementation). Our study used expert consultation and a literature review to provide additional insights and a summary of the available evidence of these underlying mechanisms and indicators that could inform future measles CFR estimations.


The anatomy of the optic chiasma and heteronymous hemianopia.
The gross anatomy of the optic nerves and chiasma has been studied, and differences in the tension in the crossed and uncrossed fibres after chiasmal displacement have been investigated. The anterior and posterior attachments of the medial and lateral fibres of the nerves have been studied. The chiasma has been dissected under low power microscopy and a three dimensional picture of it developed. Bitemporal hemianopia, as well as associated or independent hemianopic scotomata, results from stretching of the crossing fibres in the chiasma. Binasal hemianopia results from compression of the uncrossed fibres in the optic nerve or chiasma by the anterior cerebral or internal carotid arteries. The compression is effective because it is sharply localized and, probably as a result of pulsation, deeply grooves the nerve with a resulting acute distortion of fibres; it is likely that the lax lateral fibres would be less affected by a more widely spread compression. When this defect develops on top of an existing bitemporal hemianopia, it is believed that its usual cause remains the same. The crossed and uncrossed fibres of the optic chiasma differ not only anatomically in the areas of retina in which they arise but also physically. Tension is the force which occasions bitemporal hemianopia and pressure that which produces nasal field defects.


[Neurobiological correlates of suicidal behavior].
Studies in postmortem brain tissue from suicide victims show a presynaptic serotonergic deficit resulting in compensatory upregulation of postsynaptic 5-HT2 receptors in the prefrontal cortex. Reduced levels of cerebrospinal fluid (CSF) 5-hydroxyindoleacetic acid, the major serotonin metabolite, are associated with violent suicide attempts independent of psychiatric diagnosis and predict future suicide attempts and suicide completion, consistent with the notion of a biochemical trait. Neuroendocrine challenge tests, platelet studies and a polymorphism in the gene for tryptophan hydroxylase, the rate-limiting enzyme in the biosynthesis of serotonin, suggest that serotonergic activity may be reduced. This serotonergic abnormality might be related to the vulnerability or diathesis for suicidal behaviour by predisposing individuals to impulsive and autoaggressive behaviour. The hypothesis of hyperactivity of the hypothalamic pituitary adrenal axis (HPA) is supported by the postmortem findings of increased CSF corticotropin releasing hormone (CRH) concentrations and reduced CRH receptor binding sites in the frontal cortex of suicide victims (interpreted as downregulation following CRH hypersecretion) and dexamethasone nonsuppression in suicide attempters. Animal studies and in vitro experiments indicate that the HPA system modulates serotonergic activity. It is hypothesized that the serotonergic alterations potentially result from dysregulation of the HPA system. Data from epidemiological and clinical studies demonstrate that low levels of cholesterol are associated with increased suicide risk. In neuronal membranes cholesterol modulates presynaptic and postsynaptic serotonergic neurotransmission. In monkeys dietary cholesterol lowering inhibits central serotonergic activity and predisposes the animals to impulsive and aggressive behaviour. It is speculated that dysregulation of the HPA system and disordered cholesterol metabolism could enhance the serotonergic deficit, thus contributing to a neurobiological vulnerability or diathesis for impulsive and autoaggressive behaviour.


A gammaGT-AT1A receptor transgene protects renal cortical structure in AT1 receptor-deficient mice.
To understand the physiological role of angiotensin type 1 (AT(1)) receptors in the proximal tubule of the kidney, we generated a transgenic mouse line in which the major murine AT(1) receptor isoform, AT(1A), was expressed under the control of the P1 portion of the gamma-glutamyl transpeptidase (gammaGT) promoter. In transgenic mice, this promoter has been shown to confer cell-specific expression in epithelial cells of the renal proximal tubule. To avoid random integration of multiple copies of the transgene, we used gene targeting to produce mice with a single-copy transgene insertion at the hypoxanthine phosphoribosyl transferase (Hprt) locus on the X chromosome. The physiological effects of the gammaGT-AT(1A) transgene were examined on a wild-type background and in mice with targeted disruption of one or both of the murine AT(1) receptor genes (Agtr1a and Agtr1b). On all three backgrounds, gammaGT-AT(1A) transgenic mice were healthy and viable. On the wild-type background, the presence of the transgene did not affect development, blood pressure, or kidney structure. Despite relatively low levels of expression in the proximal tubule, the transgene blunted the increase in renin expression typically seen in AT(1)-deficient mice and partially rescued the kidney phenotype associated with Agtr1a(-/-)Agtr1b(-/-) mice, significantly reducing cortical cyst formation by more than threefold. However, these low levels of cell-specific expression of AT(1) receptors in the renal proximal tubule did not increase the low blood pressures or abolish sodium sensitivity, which are characteristic of AT(1) receptor-deficient mice. Although our studies do not clearly identify a role for AT(1) receptors in the proximal tubules of the kidney in blood pressure homeostasis, they support a major role for these receptors in modulating renin expression and in maintaining structural integrity of the renal cortex.


[Lymphogenic dissemination of ovarian cancer based on autopsy data].
A clinicopathological study of 152 autopsied cases of ovarian cancer was carried out. Metastases in lymph nodes ranked third (57.9%), following lesions in the peritoneum (92.1%) and greater omentum (79.6%). Metastases in lymph nodes were found in 88 cases; almost invariably, they were concomitant with implantation and hematogenous metastases. The sites of lymphogenous lesions were most frequently in paraaortal (72.7%), pelvic (47.7%) and mesenteric (27.3%) lymph nodes. No metastases in lymph collectors were detected in nearly half the cases (42.1%) of generalized malignancy. It was shown that lymphogenous dissemination does not necessarily predominate in the clinical picture even at the terminal stage of the disease.


High-resolution in vivo imaging of peripheral nerves using optical coherence tomography: a feasibility study.
Because of their complex topography, long courses, and small diameters, peripheral nerves are challenging structures for radiological diagnostics. However, imaging techniques in the area of peripheral nerve diseases have undergone unexpected development in recent decades. They include MRI and high-resolution sonography (HRS). Yet none of those imaging techniques reaches a resolution comparable to that of histological sections. Fascicles are the smallest discernable structure. Optical coherence tomography (OCT) is the first imaging technique that is able to depict a nerve's ultrastructure at micrometer resolution. In the current study, the authors present an in vivo assessment of human peripheral nerves using OCT. OCT measurement was performed in 34 patients with different peripheral nerve pathologies, i.e., nerve compression syndromes. The nerves were examined during surgery after their exposure. Only the sural nerve was twice examined ex vivo. The Thorlabs OCT systems Callisto and Ganymede were used. For intraoperative use, a hand probe was covered with a sterile foil. Different postprocessing imaging techniques were applied and evaluated. In order to highlight certain structures, five texture parameters based on gray-level co-occurrence matrices were calculated according to Haralick. The intraoperative use of OCT is easy and intuitive. Image artifacts are mainly caused by motion and the sterile foil. If the artifacts are kept at a low level, the hyporeflecting bundles of nerve fascicles and their inner parts can be displayed. In the Haralick evaluation, the second angular moment is most suitable to depict the connective tissue. OCT is a new imaging technique that has shown promise in peripheral nerve surgery for particular questions. Its resolution exceeds that provided by recent radiological possibilities such as MRI and HRS. Since its field of view is relatively small, faster acquisition times would be highly desirable and have already been demonstrated by other groups. Currently, the method resembles an optical biopsy and can be a supplement to intraoperative sonography, giving high-resolution insight into a suspect area that has been located by sonography in advance.


Circumventing surface tension: tadpoles suck bubbles to breathe air.
The surface tension of water provides a thin, elastic membrane upon which many tiny animals are adapted to live and move. We show that it may be equally important to the minute animals living beneath it by examining air-breathing mechanics in five species (three families) of anuran (frog) tadpoles. Air-breathing is essential for survival and development in most tadpoles, yet we found that all tadpoles at small body sizes were unable to break through the water's surface to access air. Nevertheless, by 3 days post-hatch and only 3 mm body length, all began to breathe air and fill the lungs. High-speed macrovideography revealed that surface tension was circumvented by a novel behaviour we call 'bubble-sucking': mouth attachment to the water's undersurface, the surface drawn into the mouth by suction, a bubble 'pinched off' within the mouth, then compressed and forced into the lungs. Growing tadpoles transitioned to air-breathing via typical surface breaching. Salamander larvae and pulmonate snails were also discovered to 'bubble-suck', and two insects used other means of circumvention, suggesting that surface tension may have a broader impact on animal phenotypes than hitherto appreciated.


Experiential History as a Tuning Parameter for Attention.
[Peer commentary on "Visual selection: usually fast and automatic; seldom slow and volitional," by J. Theeuwes]. Journal of Cognition. In his current opinion piece, Theeuwes emphasizes the role of selection history as a third source of attentional selection, beyond top-down and bottom-up mechanisms, thus challenging traditional dual-process models of attention. While we agree that selection history impacts the allocation of attention, our own work suggests that this terminology may be too restrictive, and propose the simple term history as a better reflection of the impact of learning on our selection biases. Furthermore, we propose that the role of selection/experiential history on attention may not be as a unique third source of attentional selection, but rather as a tuning parameter, allowing certain categories of item to be endowed with greater task-based or feature-driven salience in a context and history dependent manner. This conceptualization presents an alternative to abandoning dual-process models of attention altogether. Rather, we can reimagine how task-based and feature-driven processes may be controlled by past experience in a dynamic and adaptable system.


Src protein tyrosine kinase family and acute inflammatory responses.
Acute inflammatory responses are one of the major underlying mechanisms for tissue damage of multiple diseases, such as ischemia-reperfusion injury, sepsis, and acute lung injury. By use of cellular and molecular approaches and transgenic animals, Src protein tyrosine kinase (PTK) family members have been identified to be essential for the recruitment and activation of monocytes, macrophages, neutrophils, and other immune cells. Src PTKs also play a critical role in the regulation of vascular permeability and inflammatory responses in tissue cells. Importantly, animal studies have demonstrated that small chemical inhibitors for Src PTKs attenuate tissue injury and improve survival from a variety of pathological conditions related to acute inflammatory responses. Further investigation may lead to the clinical application of these inhibitors as drugs for ischemia-reperfusion injury (such as stroke and myocardial infarction), sepsis, acute lung injury, and multiple organ dysfunction syndrome.


Serum fructosamine concentration in cats with overt hyperthyroidism.
To determine the effect of hyperthyroidism on serum fructosamine concentration in cats. Cohort study. 22 cats with overt hyperthyroidism. Hyperthyroidism was diagnosed on the basis of clinical signs, detection of a palpable thyroid gland, and high total serum thyroxine (T4) concentrations. Hyperthyroid cats with abnormal serum albumin, total protein, and glucose concentrations were excluded from the study. Samples for determination of serum fructosamine concentration were obtained prior to initiating treatment. Results were compared with fructosamine concentrations in healthy cats, cats in which diabetes had recently been diagnosed, and cats with hypoproteinemia. In 6 cats, follow-up measurements were obtained 2 and 6 weeks after initiating treatment with carbimazole. Serum fructosamine concentrations ranged from 154 to 267 mumol/L (median, 198 mumol/L) and were significantly lower than values in healthy cats. Eleven (50%) of the hyperthyroid cats had serum fructosamine concentrations less than the reference range. Serum fructosamine concentrations in hyperthyroid, normoproteinemic cats did not differ from values in hypoproteinemic cats. During treatment, an increase in serum fructosamine concentration was detected. In hyperthyroid cats, concentration of serum fructosamine may be low because of accelerated protein turnover, independent of blood glucose concentration. Serum fructosamine concentrations should not be evaluated in cats with overt hyperthyroidism and diabetes mellitus. Additionally, concentration of serum fructosamine in hyperthyroid cats should not be used to differentiate between diabetes mellitus and transitory stress-related hyperglycemia.


Measurement error in estimates of sprint velocity from a laser displacement measurement device.
This study aimed to determine the measurement error associated with estimates of velocity from a laser-based device during different phases of a maximal athletic sprint. Laser-based displacement data were obtained from 10 sprinters completing a total of 89 sprints and were fitted with a fifth-order polynomial function which was differentiated to obtain instantaneous velocity data. These velocity estimates were compared against criterion high-speed video velocities at either 1, 5, 10, 30 or 50 m using a Bland-Altman analysis to assess bias and random error. Bias was highest at 1 m (+ 0.41 m/s) and tended to decrease as the measurement distance increased, with values less than + 0.10 m/s at 30 and 50 m. Random error was more consistent between distances, and reached a minimum value (±0.11 m/s) at 10 m. Laser devices offer a potentially useful time-efficient tool for assessing between-subject or between-session performance from the mid-acceleration and maximum velocity phases (i. e., at 10 m and beyond), although only differences exceeding 0.22-0.30 m/s should be considered genuine. However, laser data should not be used during the first 5 m of a sprint, and are likely of limited use for assessing within-subject variation in performance during a single session.


A phase I trial of the novel farnesyl protein transferase inhibitor, BMS-214662, in combination with paclitaxel and carboplatin in patients with advanced cancer.
This phase I study was conducted to determine the toxicities, pharmacokinetics, and pharmacodynamics of BMS-214662, a farnesyl transferase inhibitor, in combination with paclitaxel and carboplatin, in patients with advanced solid tumors. Patients with solid tumors received one of six escalating dose levels of BMS-214662 infused over 1 hour given following paclitaxel and carboplatin on the first day of a 21-day cycle. Toxicities were graded by the National Cancer Institute common toxicity criteria and recorded as maximum grade per patient for each treatment cycle. Inhibition of farnesyl transferase activity in peripheral blood mononuclear cells (PBMCs) was evaluated. Accumulation of unfarnesylated HDJ-2 in PBMCs of patients was evaluated as a marker of farnesyl transferase inhibition by BMS-214662. Thirty patients received 141 cycles of treatment through six dose levels. Dose-limiting toxicities were neutropenia, thrombocytopenia, nausea, and vomiting. There was no pharmacokinetic interaction between BMS-214662 and paclitaxel. The maximum tolerated dose was established as BMS-214662 (160 mg/m(2)), paclitaxel (225 mg/m(2)) and carboplatin (area under the curve = 6 on day 1), every 21 days. Inhibition of HDJ-2 farnesylation in PBMCs of patients was shown. One measurable partial response was observed in a patient with taxane-resistant esophageal cancer. There was partial regression of evaluable disease in two other patients (endometrial and ovarian cancer). Stable disease (> 4 cycles) occurred in eight other patients. The combination of BMS-214662 with paclitaxel and carboplatin was well tolerated, with broad activity in solid tumors. There was no correlation between dose level and accumulation of unfarnesylated HDJ-2 in PBMCs nor tumor response.


Robust three-dimensional nanotube-in-micropillar array electrodes to facilitate size independent electroporation in blood cell therapy.
Blood is an attractive carrier for plasmid and RNA-based medicine in cell therapy. Electroporation serves as a favorable delivery tool for simple operation, quick internalization, minimum cell culture involvement, and low contamination risk. However, the delivery outcome of electroporation heavily depends on the treated cells such as their type, size, and orientation to the electric field, not ideal for highly heterogeneous blood samples. Herein, a new electroporation system was developed towards effective transfection to cells in blood regardless of their large diversity. By coupling replica molding and infiltration-coating processes, we successfully configured a three-dimensional electrode comprised of a polymer micropillar array on which carbon nanotubes (CNTs) are partially embedded. During electroporation, cells sag between micropillars and deform to form a conformal contact with their top and side surfaces. The implanted CNTs not only provide a robust conductive coating for polymer micropattern but also have their protruded ends face the cell membrane vertically everywhere with maximum transmembrane potential. Regardless of their largely varied sizes and random dispersion, both individual blood cell type and whole blood samples were effectively transfected with plasmid DNA (85% after 24 h and 95% after 72 h, or 2.5-3.0 folds enhancement). High-dose RNA probes were also introduced, which regulate better the expression levels of exogenous and endogenous genes in blood cells. Besides its promising performance on non-viral delivery routes to cell-related studies and therapy, the involved new fabrication method also provides a convenient and effective way to construct flexible electronics with stable micro/nano features on the surface.


Conservative surgery for carcinoma of the cervix.
Cancer of the cervix is the second most common female cancer, with more than half a million cases worldwide occurring annually. Although screening programmes have significantly reduced the incidence and death rates in the western world, social pressures have led to the delay in childbearing, increasing numbers of cervix cancer present early in reproductive life and at an early stage. These women are often anxious to retain their fertility potential, if at all possible. Standard treatment is either radical hysterectomy or radiotherapy to the pelvis, both of which will inevitably compromise fertility, rendering future childbearing impossible. This has led to a questioning of the rationale for extensive surgery in all cases of early stage cervical cancer. The experience of reducing the radicality of surgery while not compromising its efficacy has been learnt from the current management of breast cancer. Such that this may be applied to cervical cancer. Accurate staging and measurement using magnetic resonance imaging can indicate the site and location of the tumour, allowing a radical wide local excision of early stage tumours to be carried out. This allows conservation of the corpus uteri with a potential for fertility preservation. An isthmic vaginal anastomosis maintains continuity of the vaginal canal after insertion of an isthmic cerclage. Laparoscopic pelvic lymphadenectomy completes the procedure. This technique combines aspects of the traditional radical vaginal hysterectomy plus the use of minimal access surgery to carry out the pelvic node dissection. A more invasive abdominal approach has also been described. Over 900 cases have been carried out and published, with 790 carried out vaginally (radical vaginal trachelectomy) and 116 abdominally. There have been over 300 pregnancies with 195 live births. Premature rupture of the membranes is a risk, with 10% of babies being significantly premature. Delivery is by classical caesarean section. Fertility rates are good with a low recurrence rate of 4%. Overall, radical vaginal trachelectomy seems to be a safe procedure in well-selected cases when carried out in centres with appropriate experience of radical vaginal surgery, and laparoscopic techniques. Obstetric management in high-risk feto-maternal units is necessary in view of the high risk of prematurity. This new approach preserves fertility in previously impossible situations and questions traditional management and teaching.


Cooling-promoted myogenic differentiation of murine bone marrow mesenchymal stem cells through TRPM8 activation in vitro.
TRPM8 agonist has been reported to promote osteogenic differentiation of mesenchymal stem cells (MSCs), therefore we evaluated whether cooling-induced activation of TRPM8 promotes myogenic differentiation of MSCs. We used 5-azacytidine as a myogenic differentiation inducer in murine bone marrow-derived MSCs. Addition of menthol, a TRPM8 agonist, to the differentiation induction medium significantly, increased the percentage of MyoD-positive cells, a specific marker of myogenic differentiation. We performed intracellular Ca2+ imaging experiments using fura-2 to confirm TRPM8 activation by cooling stimulation. The results confirmed that intracellular Ca2+ concentration ([Ca2+ ]i) increases due to TRPM8 activation, and TRPM8 antagonist inhibits increase in [Ca2+ ]i at medium temperatures below 19°C. We also examined the effect of cooling exposure time on myogenic differentiation of MSCs using an external cooling stimulus set at 17°C. The results showed that 60 min of cooling had an acceleratory effect on differentiation (2.18 ± 0.27 times). We observed that the TRPM8 antagonist counteracted the differentiation-promoting effect of the cooling. These results suggest that TRPM8 might modulate the multiple differentiation pathways of MSCs, and that cooling is an effective way of activating TRPM8, which regulates MSCs differentiation in vitro.


Silencing of linc00337 inhibits proliferation, cell cycle progression, migration, and invasion of colorectal cancer cells through the MEK/ERK pathway.
To explore the expression and biological functions of linc00337 in colorectal cancer (CRC), as well as its underlying mechanism. The relative expression of linc00337 in 47 cases of CRC tissues and cells was detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). The si-linc00337 interference sequences were designed and transiently transfected into CRC cells. The interference efficiency was detected via qRT-PCR. Regulatory effect of linc00337 on proliferation of CRC cells was detected via colony formation assay. Cell cycle distribution and apoptosis rate after interference in linc00337 expression were determined using flow cytometry. Moreover, the effects of linc00337 knockdown on cell migration and invasion were detected using transwell assay. At last, the effect of si-linc00337 on the MEK/ERK signaling pathway was detected using Western blotting. The results of qRT-PCR showed that among the 47 cases of CRC tissues, the expression of linc00337 was up-regulated in 40 cases. Similarly, it was highly expressed in CRC cell lines. The results of colony formation assay manifested that cell proliferation declined after interference in linc00337 expression. The results of flow cytometry and transwell assay showed that interference in linc00337 expression arrested the cell cycle in G1/G0 phase, increased the apoptosis rate, and inhibited the invasion and migration of CRC cells. According to the results of Western blotting, expressions of molecular markers in the MEK/ERK pathway after interference in linc00337 expression were significantly changed. Linc00337 is up-regulated in CRC tissues and cells. Interference in linc00337 expression can inhibit cell proliferation, migration, and invasion and promote apoptosis through the MEK/ERK pathway.


Progress and pitfalls of using isobaric mass tags for proteome profiling.
Introduction: Quantitative proteomics using mass spectrometry is performed via label-free or label-based approaches. Labeling strategies rely on the incorporation of stable heavy isotopes by metabolic, enzymatic, or chemical routes. Isobaric labeling uses chemical labels of identical masses but of different fragmentation behaviors to allow the relative quantitative comparison of peptide/protein abundances between biological samples.Areas covered: We have carried out a systematic review on the use of isobaric mass tags in proteomic research since their inception in 2003. We focused on their quantitative performances, their multiplexing evolution, as well as their broad use for relative quantification of proteins in pre-clinical models and clinical studies. Current limitations, primarily linked to the quantitative ratio distortion, as well as state-of-the-art and emerging solutions to improve their quantitative readouts are discussed.Expert opinion: The isobaric mass tag technology offers a unique opportunity to compare multiple protein samples simultaneously, allowing higher sample throughput and internal relative quantification for improved trueness and precision. Large studies can be performed when shared reference samples are introduced in multiple experiments. The technology is well suited for proteome profiling in the context of proteomic discovery studies.


Structured Curriculum to Improve Pediatric Resident Confidence and Skills in Providing Parenting Advice.
Residents receive little parenting education despite its potential to improve parenting behavior and decrease child maltreatment despite the inclusion of parenting content on board certification examinations. Teaching residents how to discuss parenting and foster positive parent-child relationships is essential to treating the whole person in osteopathic pediatric medicine. To improve pediatric and internal medicine-pediatric residents' knowledge, confidence, and skills in providing parenting advice. Four toddler parenting and discipline modules were developed. During continuity clinic, residents viewed and discussed modules with faculty. Residents completed a confidence and knowledge questionnaire before and after the curriculum, provided a self-report of use of skills learned, and completed a feasibility survey. Faculty also completed a feasibility survey. Forty-one of 61 residents (67%) participated in the study. Before participation, the median (interquartile range [IQR]) resident score for confidence in giving advice was 6.0 (4.0-7.0) (on a 10-point scale), increasing to 7.0 (6.0-8.0) for those completing 1 to 3 modules and 8.0 (8.0-9.0) for those completing 4 modules. Median (IQR) score on board-style questions was 8.0 (7.0-9.0) (on a 12-point scale) before participating in the modules and 8.5 (7.5-9.5) for those completing 1 to 3 modules and 9.0 (7.0-9.0) for those completing 4 modules after participation; the increase was not statistically significant. Nine faculty and 29 residents completed the modules and responded to the exit survey regarding feasibility and acceptability of the curriculum. On a 4-point scale (4 being excellent), sessions had an overall mean (SD) rating of 3.7 (0.5) by faculty and 3.5 (0.5) by residents. Most residents (27 [93.1%]) reported interest in more modules, and 28 residents (96.6%) reported using information learned from the modules during clinic visits. Confidence delivering parenting advice increased among residents who completed the curriculum modules. Faculty and residents reported high feasibility ratings, and residents endorsed application of skills during clinic visits and interest in more modules.


Depression of peripheral chemosensitivity by a dopaminergic mechanism in patients with obstructive sleep apnoea syndrome.
In the present study, respiratory drives to chemical stimuli and peripheral chemosensitivity were evaluated in patients with obstructive sleep apnoea (OSAS). The effects of oral administration of domperidone, a selective dopamine D2-receptor antagonist, were also examined, to study the respiratory effects of endogenous dopamine on peripheral chemoreceptors. Sixteen patients with OSAS and nine normal control subjects were studied. Respiratory responses to hypercapnia and hypoxia were measured using the rebreathing method and isocapnic progressive hypoxia method, respectively. The hypoxic withdrawal test, which measures the decrease in ventilation caused by two breaths of 100% O2 under mild hypercapnic hypoxic conditions (end-tidal oxygen and carbon dioxide tensions approximately 8.0 kPa and 5.3-6.7 kPa, respectively), was used to evaluate peripheral chemosensitivity. In the patients with OSAS, ventilatory responses to hypercapnia and hypoxia were significantly decreased compared with those of control subjects. Hypoxic withdrawal tests showed that peripheral chemosensitivity was significantly lower in patients with OSAS than in normal subjects. Hypercapnic ventilatory response and peripheral chemosensitivity were enhanced by administration of domperidone in the patients with OSAS, although no changes in either of these were observed in the control subjects. The hypoxic ventilatory response and peripheral chemosensitivity in the patients with OSAS were each significantly correlated with severity of hypoxia during sleep. These findings suggest that peripheral chemosensitivity in patients with obstructive sleep apnoea syndrome may be decreased as a result of abnormality in dopaminergic mechanisms and that the reduced chemosensitivity observed in patients with obstructive sleep apnoea syndrome may affect the severity of hypoxia during sleep.


MyCTC chip: microfluidic-based drug screen with patient-derived tumour cells from liquid biopsies.
Cancer patients with advanced disease are characterized by intrinsic challenges in predicting drug response patterns, often leading to ineffective treatment. Current clinical practice for treatment decision-making is commonly based on primary or secondary tumour biopsies, yet when disease progression accelerates, tissue biopsies are not performed on a regular basis. It is in this context that liquid biopsies may offer a unique window to uncover key vulnerabilities, providing valuable information about previously underappreciated treatment opportunities. Here, we present MyCTC chip, a novel microfluidic device enabling the isolation, culture and drug susceptibility testing of cancer cells derived from liquid biopsies. Cancer cell capture is achieved through a label-free, antigen-agnostic enrichment method, and it is followed by cultivation in dedicated conditions, allowing on-chip expansion of captured cells. Upon growth, cancer cells are then transferred to drug screen chambers located within the same device, where multiple compounds can be tested simultaneously. We demonstrate MyCTC chip performance by means of spike-in experiments with patient-derived breast circulating tumour cells, enabling >95% capture rates, as well as prospective processing of blood from breast cancer patients and ascites fluid from patients with ovarian, tubal and endometrial cancer, where sensitivity to specific chemotherapeutic agents was identified. Together, we provide evidence that MyCTC chip may be used to identify personalized drug response patterns in patients with advanced metastatic disease and with limited treatment opportunities.


Outcomes after totally minimally invasive versus hybrid and open Ivor Lewis oesophagectomy: results from the International Esodata Study Group.
Large studies comparing totally minimally invasive oesophagectomy (TMIE) with laparoscopically assisted (hybrid) oesophagectomy are lacking. Although randomized trials have compared TMIE invasive with open oesophagectomy, daily clinical practice does not always resemble the results reported in such trials. The aim of the present study was to compare complications after totally minimally invasive, hybrid and open Ivor Lewis oesophagectomy in patients with oesophageal cancer. The study was performed using data from the International Esodata Study Group registered between February 2015 and December 2019. The primary outcome was pneumonia, and secondary outcomes included the incidence and severity of anastomotic leakage, (major) complications, duration of hospital stay, escalation of care, and 90-day mortality. Data were analysed using multivariable multilevel models. Some 8640 patients were included between 2015 and 2019. Patients undergoing TMIE had a lower incidence of pneumonia than those having hybrid (10.9 versus 16.3 per cent; odds ratio (OR) 0.56, 95 per cent c.i. 0.40 to 0.80) or open (10.9 versus 17.4 per cent; OR 0.60, 0.42 to 0.84) oesophagectomy, and had a shorter hospital stay (median 10 (i.q.r. 8-16) days versus 14 (11-19) days (P = 0.041) and 11 (9-16) days (P = 0.027) respectively). The rate of anastomotic leakage was higher after TMIE than hybrid (15.1 versus 10.7 per cent; OR 1.47, 1.01 to 2.13) or open (15.1 versus 7.3 per cent; OR 1.73, 1.26 to 2.38) procedures. Compared with hybrid and open Ivor Lewis oesophagectomy, TMIE resulted in a lower pneumonia rate, a shorter duration of hospital stay, but higher anastomotic leakage rates. Therefore, no clear advantage was seen for either TMIE, hybrid or open Ivor Lewis oesophagectomy when performed in daily clinical practice.


CPAP after endoscopic procedures as add-on therapy for the treatment of tracheal stenosis: a case series.
Tracheal stenosis represents a possible complication in intubated or tracheotomised patients. Tracheal resection is currently the gold standard for the treatment of complex stenosis while granulomas and simple stenosis (e.g., web-like) are often treated by endoscopic procedures, which do not consistently give satisfactory long-term results, due to frequent relapses. Administering continuous positive airway pressure (CPAP) after endoscopic procedures might represent a new add-on option for the treatment of this complication. In this case series are presented two patients with tracheal stenosis showed after the removal of tracheostomy tube, both treated with CPAP. The results were straightforward: CPAP treatment helped to keep stable the tracheal lumen, without adverse effects. No further endoscopic dilations were necessary thereafter, with a likely positive impact on patients' quality of life and on health expenditure.


Preparation for management: an exploratory study to ascertain perceptions of F grade nurses and clinical nurse managers.
This paper reviews relevant literature and discusses the results of a small study designed to investigate how nurses are responding to the changes in the structure of the NHS which have reduced their role in management. The findings suggest that nurses who are currently at F grade level and their clinical nurse managers (CNM) are aware of the need for nurses seeking career progression to participate in education and training programmes related to nursing and management, particularly those which provide recognized qualifications, and that many are currently striving to achieve this. The findings also suggest that nurses want support to enable them to undertake further study, but that due to increasing pressure of work it is becoming more and more difficult for CNMs to provide the level of support they consider helpful. Nurses and CNMs urge those developing courses in management to ensure that courses are flexible to enable varying types of participation and tailoring to individual needs.


Synthesis and insecticidal activities of novel anthranilic diamides containing acylthiourea and acylurea.
Two series of anthranilic diamides containing acylthiourea and acylurea linkers were designed and synthesized, with changed length and flexibility of the linkers to compare to known anthranilic diamide insecticides. In total, 26 novel compounds were synthesized, and all compounds were characterized by (1)H nuclear magnetic resonance and high-resolution mass spectrometry. Their insecticidal activities against oriental armyworm (Mythimna separata), mosquito larvae (Culex pipiens pallens), and diamondback moth (Plutella xylostella) were evaluated. The larvicidal activities against oriental armyworm indicated that the introduction of acylthiourea into some structures could retain their insecticidal activity; 8 of the 15 compounds (13a-13e, 14a-14e, and 15a-15e) exhibited 100% larvicidal activity at 10 mg/L. However, the introduction of acylurea decreased the insecticidal activity; only 3 of the 11 compounds (17a-17k) exhibited 100% larvicidal activity at 200 mg/L. The whole-cell patch-clamp technique indicated that compound 13b and chlorantraniliprole exhibited similar effects on the voltage-gated calcium channel. The calcium-imaging technique was also applied to investigate the effects of compounds 13b and 15a on the intracellular calcium ion concentration ([Ca(2+)](i)), which indicated that they released stored calcium ions from endoplasmic reticulum. Experimental results denoted that several new compounds are potential activators of the insect ryanodine receptor (RyR).


Cross-sectional imaging of common and unusual complications after endoscopic retrograde cholangiopancreatography.
Endoscopic retrograde cholangiopancreatography (ERCP) is currently a primarily therapeutic procedure that is extensively employed to treat several biliopancreatic disorders. Although widely considered a safe procedure, ERCP is associated with a non-negligible morbidity and occasional mortality. Due to the number and complexity of operative ERCPs performed, radiologists are increasingly faced with urgent requests for investigation of suspected post-procedural complications, which often have similar clinical and laboratory manifestations. This pictorial essay reviews the usual post-procedural CT findings, the clinical features and imaging appearances of common and unusual post-ERCP occurrences including interstitial oedematous and necrotising acute pancreatitis, haemorrhages, retroperitoneal and intraperitoneal duodenal perforations, infections and stent-related complications. Emphasis is placed on the pivotal role of multidetector CT, which is warranted after complex or prolonged ERCP procedures as it represents the most effective modality to detect and grade ERCP-related complications and to monitor nonsurgically treated patients. Timely diagnosis and optimal management require a combination of clinical and laboratory data with imaging appearances; therefore, this article aims to provide an increased familiarity with interpretation of early post-ERCP studies, particularly to triage those occurrences that require interventional or surgical treatment. In selected patients MRI allows imaging pancreatitis and abnormal collections without the use of ionising radiation. Teaching Points • Endoscopic retrograde cholangiopancreatography (ERCP) allows treating many biliopancreatic disorders.• Due to the number and complexity of procedures, post-ERCP complications are increasingly encountered.• Main complications include acute pancreatitis, haemorrhages, duodenal perforation and infections.• Diagnosis and management of complications rely on combined clinical, laboratory and imaging data.• Multidetector CT is most effective to diagnose, categorise and monitor post-ERCP complications.


Synthesis, physicochemical, rheological and in-vitro characterization of double-crosslinked hyaluronic acid hydrogels containing dexamethasone and PLGA/dexamethasone nanoparticles as hybrid systems for specific medical applications.
Injectable hydrogels and biodegradable nanoparticles are using in tissue engineering applications and drug delivery systems. To improve physiochemical properties of biomaterials and to develop their applications, hybrid systems consist of hydrogels, and biodegradable nanoparticles were synthesized. In this study, hybrid systems based on double crosslinked hyaluronic acid and PLGA/Dexamethasone sodium phosphate (PLGADEX) nanoparticles are designed and synthesized in several steps. At the first step, poly(l-lactide-co-glycolide) (PLGA) in a ratio of LLA:GA = 85:15 mol% was synthesized via ring-opening polymerization. Then, PLGADEX nanoparticles were synthesized in different ratios using the partially modified emulsification-diffusion method and fully characterized, and desirable nanoparticle was selected (PLGADEX20). At the second step, a double cross-linked hyaluronic acid (XHA) was prepared by mixing various ratios of amino-hyaluronic acid and aldehyde-hyaluronic acid in the presence of genipin. Finally, by mixing of various ratios of PLGADEX20 and Dexamethasone sodium phosphate (DEX) with different ratios of XHA, hybrid systems were prepared. Based on the characterization of hybrid samples and the release studies, hydrogels containing nanoparticles showed a controlled drug release, while the best sample with 3% of optimized nanoparticle was chosen. According to physiochemical and biological properties, these hybrid systems can be good candidates for anti-adhesion barriers, wound dressings, and novel drug delivery systems.


Elongation of fatty acids by microsomal fractions from the brain of the developing rat.
Elongation of fatty acids by microsomal fractions obtained from rat brain was measured by the incorporation of [2-14C]malonyl-CoA into fatty in the presence of palmitoyl-CoA or stearoyl-CoA. 2. Soluble and microsomal fractions were prepared from 21-day-old rats; density gradient centrifugation demonstrated that the stearoyl-CoA elongation system was localized in the microsomal fraction whereas fatty acid biosynthesis de novo from acetyl-CoA occurred in the soluble fraction. The residual activity de novo in the microsomal fraction was attributed to minor contamination by the soluble fraction. 3. The optimum concentration of [2-14C]malonyl-CoA for elongation of fatty acids was 25 mum for palmitoyl-CoA or stearoyl-CoA, and the corresponding optimum concentrations for the two primer acyl-CoA esters were 8.0 and 7.2 muM respectively. 4. Nadph was the preferred cofactor for fatty acid formation from palmitoyl-CoA or stearoyl-CoA, although NADH could partially replace it. 5. The stearoyl-CoA elongation system required a potassium phosphate buffer concentration of 0.075M for maximum activity; CoA (1 MUM) inhibited this elongation system by approx. 30%. 6. The fatty acids formed from malonyl-CoA and palmitoyl-CoA had a predominant chain length of C18 whereas stearoyl-CoA elongation resulted in an even distribution of fatty acids with chain lengths of C20, C22 and C24. 7. The products of stearoyl-CoA elongation were identified as primarily unesterified fatty acids. 8. The developmental pattern of fatty acid biosynthesis by rat brain microsomal preparations was studied and both the palmitoyl-CoA and stearoyl-CoA elongation systems showed large increases in activity between days 10 and 18 after birth.


[Perineal ectopic testis].
Three further cases of perineal ectopic testis are reported and the published literature reviewed. Etiopathogenicity is discussed and emphasis placed on the need for perineal exploration in all cases of absence of testes from the scrotum. The most effective route of approach for repair is an inguinal one allowing replacement of the testis into the corresponding hemiscrotum without difficulty. Because of the histopathologic features involved, prognosis is better than that of cryptorchidism.


Prelude to corneal tissue engineering - gaining control of collagen organization.
By most standard engineering practice principles, it is premature to credibly discuss the "engineering" of a human cornea. A professional design engineer would assert that we still do not know what a cornea is (and correctly so), therefore we cannot possibly build one. The proof resides in the fact that there are no clinically viable corneas based on classical tissue engineering methods available. This is possibly because tissue engineering in the classical sense (seeding a degradable scaffolding with a population synthetically active cells) does not produce conditions which support the generation of organized tissue. Alternative approaches to the problem are in their infancy and include the methods which attempt to recapitulate development or to produce corneal stromal analogs de novo which require minimal remodeling. Nonetheless, tissue engineering efforts, which have been focused on producing the fundamental functional component of a cornea (organized alternating arrays of collagen or "lamellae"), may have already provided valuable new insights and tools relevant to development, growth, remodeling and pathologies associated with connective tissue in general. This is because engineers ask a fundamentally different question (How can that be done?) than do biological scientists (How is that done?). The difference in inquiry has prompted us to closely examine (and to mimic) development as well as investigate collagen physicochemical behavior so that we may exert control over organization both in cell culture (in vitro) and on the benchtop (de novo). Our initial results indicate that reproducing corneal stroma-like local and long-range organization of collagen may be simpler than we anticipated while controlling spacing and fibril morphology remains difficult, but perhaps not impossible in the (reasonably) near term.